Intelligent manufacturing systems that enable real-time product releases will become commonplace in the not-too-distant future, thanks to advances in process monitoring and analytical techniques, according to researchers from CSL Innovation and Philipps-Universität Marburg.
Many of those new and notable techniques and technologies were highlighted in a recent paper by Pavithra Sathiyapriyan, first author, PhD student at Philipps-Universität Marburg and researcher at CSL Innovation; Uwe Kalina, PhD, senior author and researcher at CSL; and colleagues.
As biopharmaceutical manufacturing shifts towards continuous manufacturing, “Implementation of process analytical technology (PAT) in downstream processing has been somewhat limited,” the team acknowledges, despite being introduced in 2004. Reasons include the complexity of therapeutic proteins, the heterogeneity of protein structures in any mixture, and the need for immediate responses. Addressing those points requires an integrated approach involving multiple in-line, on-line, and at-line techniques and technologies, they stress.
Some of the most exciting advances are occurring in spectroscopy, reducing the challenges of rapid signal saturation and the need for extensive sample preparation.
For example, combining mid-infrared spectroscopy (MIR) with Fourier transform infrared spectroscopy (FTIR) reduces the constraints and “makes it useful in measuring product concentrations in complex matrices,” they report. Attenuated total reflectance (ATR) further mitigates MIR’s limitations by probing only a thin sample layer, thereby “preventing sample saturation by interfering molecules such as water.” By monitoring multiple parameters simultaneously in real time, ATR-FTIR—when combined with certain algorithms—enables real-time prediction and control, Sathiyapriyan and colleagues note.
Additionally, they continue, “Surface-enhanced infrared absorption spectroscopy (SEIRA) and tip-enhanced IR methods can increase measurement sensitivity by several fold,” and make in situ monitoring applications feasible.
Near-infrared spectroscopy (NIR) penetrates more deeply than MIR and is useful in high-throughput analysis with minimal sample preparation. Fourier transform techniques and NIR have monitored column loading, and researchers in India used NIR to simultaneously monitor protein and excipient concentrations in antibody formulations. Results were delivered in real time.
Ramen spectroscopy—which measures polarity changes rather than the diploe changes recorded by infrared spectroscopy—can identify a sample’s molecule composition and structural information. Limitations, however, include sensitivity to sample temperature and fluorescence interference.
Recent advances, such as red or deep-red laser wavelengths, time-resolved Raman, and shifted excitation Ramen difference spectroscopy (SERDS), reduce fluorescence interference. Advantages of other variants—such as resonance Raman, surface-enhanced Raman (SERS), surface-enhanced resonance Raman (SERRS), and tip-enhanced Taman (TERS)—include increased signal strength, lowered detection limits, and reduced the sample volume needed.
UV-vis spectroscopy, another new addition to PAT, measures electron transitions and therefore mainly detects chromophores—”regions where electron excitation is more likely.” One study showed this method, combined with partial least squares regression modeling, accurately predicted monoclonal antibody concentrations during the loading phase of Protein A chromatography.
Variable pathlength slope (VPE) instruments, which measure absorbance automatically at a specific wavelength, are making it possible now for UV-vis methods to accurately measure even high-concentration, undiluted proteins, the scientists note.
Each of these techniques—and many others—are enhancing the throughput, sensitivity, and measurement speed needed for commercial-scale PAT. They still lack the specificity needed to be practical, the authors admit, but, as they say, “Advancements are continually evolving. The emphasis should extend beyond individual techniques to optimizing the integration and effective utilization of acquired data.”
The post Spectroscopy Advances Support PAT in Downstream Processing Operations appeared first on GEN - Genetic Engineering and Biotechnology News.
Many of those new and notable techniques and technologies were highlighted in a recent paper by Pavithra Sathiyapriyan, first author, PhD student at Philipps-Universität Marburg and researcher at CSL Innovation; Uwe Kalina, PhD, senior author and researcher at CSL; and colleagues.
As biopharmaceutical manufacturing shifts towards continuous manufacturing, “Implementation of process analytical technology (PAT) in downstream processing has been somewhat limited,” the team acknowledges, despite being introduced in 2004. Reasons include the complexity of therapeutic proteins, the heterogeneity of protein structures in any mixture, and the need for immediate responses. Addressing those points requires an integrated approach involving multiple in-line, on-line, and at-line techniques and technologies, they stress.
Spectroscopy moves forward
Some of the most exciting advances are occurring in spectroscopy, reducing the challenges of rapid signal saturation and the need for extensive sample preparation.
For example, combining mid-infrared spectroscopy (MIR) with Fourier transform infrared spectroscopy (FTIR) reduces the constraints and “makes it useful in measuring product concentrations in complex matrices,” they report. Attenuated total reflectance (ATR) further mitigates MIR’s limitations by probing only a thin sample layer, thereby “preventing sample saturation by interfering molecules such as water.” By monitoring multiple parameters simultaneously in real time, ATR-FTIR—when combined with certain algorithms—enables real-time prediction and control, Sathiyapriyan and colleagues note.
Additionally, they continue, “Surface-enhanced infrared absorption spectroscopy (SEIRA) and tip-enhanced IR methods can increase measurement sensitivity by several fold,” and make in situ monitoring applications feasible.
Near-infrared spectroscopy (NIR) penetrates more deeply than MIR and is useful in high-throughput analysis with minimal sample preparation. Fourier transform techniques and NIR have monitored column loading, and researchers in India used NIR to simultaneously monitor protein and excipient concentrations in antibody formulations. Results were delivered in real time.
Ramen spectroscopy—which measures polarity changes rather than the diploe changes recorded by infrared spectroscopy—can identify a sample’s molecule composition and structural information. Limitations, however, include sensitivity to sample temperature and fluorescence interference.
Recent advances, such as red or deep-red laser wavelengths, time-resolved Raman, and shifted excitation Ramen difference spectroscopy (SERDS), reduce fluorescence interference. Advantages of other variants—such as resonance Raman, surface-enhanced Raman (SERS), surface-enhanced resonance Raman (SERRS), and tip-enhanced Taman (TERS)—include increased signal strength, lowered detection limits, and reduced the sample volume needed.
UV-vis spectroscopy, another new addition to PAT, measures electron transitions and therefore mainly detects chromophores—”regions where electron excitation is more likely.” One study showed this method, combined with partial least squares regression modeling, accurately predicted monoclonal antibody concentrations during the loading phase of Protein A chromatography.
Variable pathlength slope (VPE) instruments, which measure absorbance automatically at a specific wavelength, are making it possible now for UV-vis methods to accurately measure even high-concentration, undiluted proteins, the scientists note.
Each of these techniques—and many others—are enhancing the throughput, sensitivity, and measurement speed needed for commercial-scale PAT. They still lack the specificity needed to be practical, the authors admit, but, as they say, “Advancements are continually evolving. The emphasis should extend beyond individual techniques to optimizing the integration and effective utilization of acquired data.”
The post Spectroscopy Advances Support PAT in Downstream Processing Operations appeared first on GEN - Genetic Engineering and Biotechnology News.